Specific killing of latently HIV-1-infected cells after provirus reactivation

原病毒重新激活后特异性杀死潜伏 HIV-1 感染的细胞

• 批准号: 9040568
• 负责人: Qigui Q Yu
• 金额: $ 38.64万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9040568
• 关键词: Anti-Retroviral Agents; Antibodies; Antibody Response; Autologous; Biological Assay; CD4 Positive T Lymphocytes; Cell Line; Cells; Complement; Complement Activation; Cytolysis; Detection; Diagnostic; Failure; Gene Expression; Goals; Grant; HIV-1; Highly Active Antiretroviral Therapy; Human; Immune response; Immunity; In Vitro; Individual; Infection; Interleukin-7; Life; Measurable; Mediating; Medical; Memory; Modeling; Morbidity - disease rate; Patients; Pharmaceutical Preparations; Phase; Production; Proliferating; Proviruses; Public Health; Reporting; Research; Residual state; Rest; Serum; Surface; T-Lymphocyte; Testing; Vaccines; Valproic Acid; Viral; Viral Cytopathogenic Effect; Viral Proteins; Viremia; Virion; Virus; Vorinostat; antiretroviral therapy; cell type; effective therapy; experience; global health; improved; in vivo; inhibitor/antagonist; innovation; killings; member; mortality; novel strategies; pandemic disease; prostratin; purge

PROJECT SUMMARY/ABSTRACT The HIV-1 pandemic has claimed over 20 million lives, with 38.6 million people worldwide currently infected, and will continue to be a significant global health problem as there is no vaccine available. Currently, the only effective treatment available to HIV-1 infection is HAART (highly active antiretroviral therapy), which has led to a profound reduction in HIV-1-related morbidity and mortality. However, HAART fails to eliminate the virus in vivo, mainly due to the persistent existence of long-lived latently-infected cells harboring replication-competent proviruses. Efforts to purge these infected cells have focused on reactivation of the proviruses. It is presumed that these infected cells will be killed after reactivation of virus gene expression by viral cytopathic effects (CPEs), host immune responses or both. Several stimulants including IL-7, valproic acid (VPA), suberoylanilide hydroxamic acid (SAHA) and prostratin have been explored to force activation of proviruses in latently-infected resting CD4+ T cells that constitute the major reservoir of HIV-1 in vivo. However, treatment with IL-7 or VPA in patients on HAART has failed to reduce HIV-1 latency, suggesting that these agents alone are not sufficient to induce killing of latently-infected cells. We propose to develop an “activation-killing” approach by combining provirus stimulants with an agent that is able to allow anti-HIV-1 immunity naturally mounted in patients during HIV-1 infection to specifically kill latently-infected cells after provirus reactivation. Studies have shown that both HIV-1-infected cells and virions use their surface regulators of complement activation (RCA) to resist antibody- dependent complement-mediated lysis (ADCML), which explains why vigorous and sustained anti-HIV-1 envelope (Env) antibody (Ab) responses in almost all infected individuals fail to control HIV-1 infection. We and others have reported that blocking human CD59 (hCD59), a key member of RCA, renders both HIV-1-infected cells and virions sensitive to ADCML. We therefore hypothesize that addition of an hCD59 inhibitor to SAHA, prostratin or both will enable anti-HIV-1 Env Abs naturally present in HIV-1-infected individuals to trigger ADCML of latently-infected T cells after provirus reactivation. The provirus stimulants will activate proviruses to express viral proteins on the surface of latently-infected cells that will be killed by ADCML if hCD59 is inhibited. We will use the human T cell line ACH2, a well-characterized model of HIV-1-latency, to develop such an approach (Aim 1, R21 Phase). We will verify whether the approach is able to trigger ADCML of the primary CD4+ T cells latently infected with HIV-1 in vitro (Aim 2, R33 Phase). We will then determine if this approach can allow anti-HIV-1 Abs naturally present in the sera of infected patients to trigger autologous ADCML of their own latently-infected resting CD4+ T cells (Aim 3, R33 Phase). Our project is innovative and significant because we aim to develop a novel approach for purging the persistent reservoir of HIV-1 in patients on HAART. This approach may simultaneously target various latently infected cells and residual viremia, potentially leading to a broader impact and improved efficacy against the various persistent HIV-1 reservoirs.

项目总结/摘要 HIV-1大流行病已夺去2 000多万人的生命，目前全世界有3 860万人受到感染， 并且由于没有可用的疫苗，它将继续成为一个重大的全球健康问题。目前，唯一 HIV-1感染的有效治疗方法是HAART（高效抗逆转录病毒疗法）， 与艾滋病毒1有关的发病率和死亡率大幅下降。然而，HAART未能消除病毒， 体内，主要是由于长期存在的潜伏感染的细胞窝藏复制能力 前病毒清除这些受感染细胞的努力集中在重新激活前病毒上。据推测 这些被感染的细胞在病毒基因表达被病毒致细胞病变效应重新激活后将被杀死 （CPE）、宿主免疫反应或两者。几种兴奋剂，包括IL-7、丙戊酸（VPA）、辛二酰苯胺 异羟肟酸（SAHA）和prostratin已被探索，以迫使激活前病毒在潜伏感染 静止的CD 4 + T细胞，构成体内HIV-1的主要储存库。然而，IL-7或VPA治疗 接受HAART治疗的患者未能减少HIV-1潜伏期，这表明单独使用这些药物不足以 诱导杀死潜伏感染细胞。我们建议开发一种“激活-杀死”的方法， 前病毒刺激剂与能够使抗HIV-1免疫在患者体内自然建立的试剂， HIV-1感染在前病毒再激活后特异性杀死潜伏感染的细胞。研究表明，两者 HIV-1感染的细胞和病毒体使用其补体激活（RCA）的表面调节剂来抵抗抗体- 依赖补体介导的裂解（ADCML），这解释了为什么强有力的和持续的抗HIV-1 包膜（Env）抗体（Ab）反应在几乎所有感染的个体不能控制HIV-1感染。我们和 其他人报道，阻断RCA的关键成员人CD 59（hCD 59）， 对ADCML敏感的细胞和病毒体。因此，我们假设在SAHA中加入hCD 59抑制剂， prostratin或两者将使天然存在于HIV-1感染个体中的抗HIV-1 Env Ab能够触发 前病毒再激活后潜伏感染T细胞的ADCML。前病毒刺激剂将激活前病毒， 在潜伏感染细胞的表面上表达病毒蛋白，如果hCD 59被抑制，这些病毒蛋白将被ADCML杀死。 我们将使用人类T细胞系ACH 2，一种充分表征的HIV-1潜伏期模型，来开发这样一种新的免疫抑制剂。 方法（目标1，R21阶段）。我们将验证该方法是否能够触发主设备的ADCML 体外潜伏感染HIV-1的CD 4 + T细胞（Aim 2，R33阶段）。然后我们将确定这种方法是否 可以使感染患者血清中天然存在的抗HIV-1抗体触发其自身的ADCML， 自身潜伏感染的静息CD 4 + T细胞（Aim 3，R33阶段）。我们的项目具有创新性和重大意义 因为我们的目标是开发一种新的方法来清除患者体内持续存在的HIV-1， HAART。这种方法可以同时靶向各种潜伏感染的细胞和残留的病毒血症， 可能导致更广泛的影响和提高对各种持续HIV-1储库的功效。